Effective dosages of an adenoviral-based biological delivery and expression system for use in the treatment of osteoarthritis in humans, and compositions comprising the same

Abstract

The disclosure relates to pharmaceutical compositions and methods of using pharmaceutical compositions comprising effective dosages of an adenoviral-based biological delivery and expression system for use in the treatment or prevention of osteoarthritis in human or mammalian joints by long-term inducible gene expression of human or mammalian interleukin-1 receptor antagonist (IL-1Ra) in synovial cells, comprising a helper-dependent adenoviral vector containing a nucleic acid sequence encoding for human or mammalian interleukin-1 receptor antagonist (IL-1Ra), left and right inverted terminal repeats (L ITR and R ITR), the adenoviral packaging signal and non-viral, non-coding stuffer nucleic acid sequences, wherein the expression of the human or mammalian interleukin-1 receptor antagonist (IL-1Ra) gene within synovial cells is regulated by an inflammation-sensitive promoter.

What is claimed: 
     
         1 . A pharmaceutical composition comprising an adenoviral-based biological delivery and expression system for the treatment of osteoarthritis or an osteoarthritic condition in a human joint or for the prevention of such conditions in a human identified to be at risk of developing osteoarthritis or an osteoarthritic condition,
 wherein the adenoviral-based biological delivery and expression system comprises genome copies (GC) of a helper-dependent adenoviral vector comprising a nucleic acid sequence encoding a human interleukin-1 receptor antagonist (IL-1Ra) protein, left and right inverted terminal repeats, an adenoviral packaging signal and non-viral, and non-coding stuffer nucleic acid sequences,   wherein the expression of the human IL-1Ra gene is regulated by a NF-κB inducible promoter, which is located upstream of the reading frame of the nucleic acid sequence encoding the human IL-1Ra protein,   wherein the nucleic acid sequence of the adenoviral-based biological delivery and expression system comprising the promoter, the nucleic acid sequence encoding the IL-1Ra, the left and the right inverted terminal repeats, the adenoviral packaging signal and the non-viral, non-coding stuffer nucleic acid sequences is at least 95% homologous to the nucleic acid sequence of SEQ ID NO: 7, and   wherein adenoviral-based biological delivery and expression system comprises 1.4×10 8  to 1.4×10 12  GC of the helper-dependent adenoviral vector per milliliter (GC per ml).   
     
     
         2 . The pharmaceutical composition according to  claim 1 , wherein the nucleic acid sequence of the adenoviral-based biological delivery and expression system comprising the promoter, the nucleic acid sequence encoding the IL-1Ra, the left and the right inverted terminal repeats, the adenoviral packaging signal and the non-viral, non-coding stuffer nucleic acid sequences is at least 99% homologous to the nucleic acid sequence of SEQ ID NO: 7. 
     
     
         3 . The pharmaceutical composition according to  claim 1 , wherein the nucleic acid sequence of the adenoviral-based biological delivery and expression system comprising the promoter, the nucleic acid sequence encoding the IL-1Ra, the left and the right inverted terminal repeats, the adenoviral packaging signal and the non-viral, non-coding stuffer nucleic acid sequences comprises the nucleic acid sequence of SEQ ID NO: 7. 
     
     
         4 . The pharmaceutical composition according to  claim 1 , wherein the IL-1Ra in the nucleic acid sequence of the adenoviral-based biological delivery and expression system comprises the nucleic acid of SEQ ID NO 4. 
     
     
         5 . The pharmaceutical composition according to  claim 4 , wherein the nucleic acid according to SEQ ID NO: 4 expresses a human IL-1Ra protein of amino acid sequence that is at least 95% homologous to SEQ ID NO: 6. 
     
     
         6 . The pharmaceutical composition according to  claim 1 , wherein the adenoviral-based biological delivery and expression system comprises:
 a) 1.4×10 9  to 1.4×10 12 ;   b) 1.4×10 9  to 1.4×10 11 ; or   c) 1.4×10 9  to 1.4×10 10 , GC per ml.   
     
     
         7 . The pharmaceutical composition according to  claim 6 , wherein the adenoviral-based biological delivery and expression system comprises 1.4×10 9  to 5.6×10 9  GC per ml. 
     
     
         8 . The pharmaceutical composition according to  claim 6 , wherein the adenoviral-based biological delivery and expression system comprises 1.4×10 10  to 5.6×10 10  GC per ml. 
     
     
         9 . The pharmaceutical composition according to  claim 6 , wherein the adenoviral-based biological delivery and expression system comprises 1.4×10 11  to 5.6×10 11  GC per ml. 
     
     
         10 . The pharmaceutical composition according to  claim 1 , wherein the adenoviral-based biological delivery and expression system comprises a dose volume of up to 5 ml. 
     
     
         11 . The pharmaceutical composition according to  claim 7 , wherein the adenoviral-based biological delivery and expression system comprises a total dose of 7×10 9  to 2.8×10 10  GC. 
     
     
         12 . The pharmaceutical composition according to  claim 8 , wherein the adenoviral-based biological delivery and expression system comprises a total dose of 7×10 10  to 2.8×10 11  GC. 
     
     
         13 . The pharmaceutical composition according to  claim 9 , wherein the adenoviral-based biological delivery and expression system comprises a total dose of 7×10 11  to 2.8×10 12  GC. 
     
     
         14 . The pharmaceutical composition according to  claim 1 , wherein the pharmaceutical composition is formulated for intra-tendinous, intra-muscular, intra-articular, or sub-acromial injection to the human joint. 
     
     
         15 . The pharmaceutical composition according to  claim 1 , wherein the pharmaceutical composition is formulated for intra-articular injection into the human joint. 
     
     
         16 . A method of infecting joint cells of one or more osteoarthritis-affected joints of a human suffering from osteoarthritis or an osteoarthritic condition with an adenoviral-based biological delivery and expression system, wherein the method comprises the steps of:
 a) infecting the joint cells of the osteoarthritis-affected joint of the human in need thereof with the pharmaceutical composition comprising an adenoviral-based biological delivery and expression system of  claim 1 ; and   b) expressing IL-1Ra in the target area within the osteoarthritis-affected joint.   
     
     
         17 . The method according to  claim 16 , wherein the joint cells are infected once with the adenoviral-based biological delivery and expression system. 
     
     
         18 . The method according to  claim 16 , wherein joint cells are infected two or more times with the adenoviral-based biological delivery and expression system. 
     
     
         19 . The method according to  claim 18 , wherein, when the joint cells are infected two or more times with an adenoviral-based biological delivery and expression system, each infection comprises a different number of genome copies of the helper-dependent adenoviral vector. 
     
     
         20 . The method according to  claim 18 , wherein, when the joint cells are infected two or more times with an adenoviral-based biological delivery and expression system, each infection comprises the same number of genome copies of the helper-dependent adenoviral vector. 
     
     
         21 . The method according to  claim 18 , wherein, when the joint cells are infected two or more times with an adenoviral-based biological delivery and expression system, each infection is done in the same osteoarthritis-affected joint of the human. 
     
     
         22 . The method according to  claim 18 , wherein, when the joint cells are infected two or more times with an adenoviral-based biological delivery and expression system, every second and subsequent infection is done in an osteoarthritis-affected joint of the human that is different than the osteoarthritis-affected joint in which the previous infection was done. 
     
     
         23 . The method according to  claim 16 , wherein the infecting of the joint cells comprises intra-articular, intra-tendinous, intra-muscular, or sub-acromial injection. 
     
     
         24 . The method of  claim 16 , further comprises the step of:
 c) monitoring the treatment or progress of osteoarthritis or an osteoarthritic condition in the osteoarthritis-affected joint following the expression of the IL-1Ra in (b).   
     
     
         25 . The method of  claim 24 , further comprises the steps of:
 (d) continuing to administer the amount of the adenoviral-based biological delivery and expression system to the osteoarthritis-affected joint of (a), if monitoring of (c) shows that the osteoarthritis or an osteoarthritic condition in the human joint is not managed or treated; or   (e) further adjusting the number of genome copies of the helper-dependent adenoviral vector in the amount of the adenoviral-based biological delivery and expression system and administering to the osteoarthritis-affected joint of (a), if monitoring of (c) shows that the osteoarthritis or an osteoarthritic condition in the human joint has progressed.   
     
     
         26 . An adenoviral-based biological delivery and expression system for treatment of osteoarthritis or an osteoarthritic condition in a human joint or for the prevention of such conditions in a human identified to be at risk of developing osteoarthritis or an osteoarthritic condition,
 wherein the adenoviral-based biological delivery and expression system comprises genome copies (GC) of a helper-dependent adenoviral vector comprising a nucleic acid sequence encoding a human interleukin-1 receptor antagonist (IL-1Ra), left and right inverted terminal repeats, an adenoviral packaging signal and non-viral, and non-coding stuffer nucleic acid sequences,   wherein the expression of the human IL-1Ra gene is regulated by a NF-κB inducible promoter, which is located upstream of the reading frame of the nucleic acid sequence encoding the human IL-1Ra,   wherein the nucleic acid sequence of the adenoviral-based biological delivery and expression system comprising the promoter, the nucleic acid sequence encoding the IL-1Ra, the left and the right inverted terminal repeats, the adenoviral packaging signal and the non-viral, non-coding stuffer nucleic acid sequences is at least 95% homologous to the nucleic acid sequence of SEQ ID NO: 7,   wherein the adenoviral-based biological delivery and expression system is isolated from a host cell that is infected with the helper-dependent adenoviral vector and a helper virus,   wherein the adenoviral-based biological delivery and expression system comprises:   a) 1.4×10 8  to 1.4×10 12  GC of the helper-dependent adenoviral vector per milliliter (GC per ml) of synovial fluid in a human joint;   b) less than 15% helper virus particles;   c) less than 10% empty capsids;   d) not more than 100 μg/ml of host cell protein;   e) not more than 20 ng/ml of host cell nucleic acid;   f) not more than 35EU/ml of endotoxin; and   g) a Viral Particle to Infectious Unit Ratio of ≤than 300GC/TCID 50 .